重组腺病毒IκBαM抑制高糖诱导的血管内皮细胞炎症因子的异常分泌

Recombinant Adenovirus IκBαM IκBαhiabit the IκBαcreased Expression of Inflammatory Factors Mediated by High Doses of Glucose in Vascular Endothelial Cells

构建携有核因子κB抑制物IκBα的突变体IκBαM的重组腺病毒(AdIκBαM),并感染ECV304血管内皮细胞,采用Westem blot、电泳迁移率变动分析和逆转录-聚合酶链反应等方法研究核因子κB在高浓度葡萄糖刺激的ECV304细胞分泌功能中所起的作用以及阻断其活性对内皮细胞分泌功能的影响。结果发现,高糖能诱导ECV304细胞IκBα的降解和核因子κB的激活,同时上调炎症因子细胞间粘附分子1、单核细胞趋化蛋白1和内皮素1 mRNA表达水平,而感染AdIκBαM的ECV304/IκBαM细胞则能拮抗高糖所致的IκBα降解与核因子κB激活,同时下调这些炎症因子的异常分泌水平。结果提示,核因子κB的激活在高糖所致的内皮细胞分泌功能改变中起中轴作用,抑制核因子κB的活化,有助于改善血管内皮细胞功能。

Aim To explore the role of nuclear factor-κB (NF-κB) in high doses of glucose (HG) induced impairment of ECV304 cells, a vascular endothelial cell line. Methods Recombinant adenovirus mediated nuclear factor-KB supper-repres sor IκBαM with mutant IκBα was constructed. Western blot, electrophoretic mobility shift assay (EMSA) and reverse tran scriptase-polymerase chain reaction(RT-PCR) assay were applied in this study. Results HG induced IκBα degradation and nuclear factor-KB activation were found in ECV304 cells but not in ECV304/IκBαM cells infected with IκBαM recombinant adenovirus . The expression of inflammatory factors such as monocyte chemotactic protein-1 (MCP-1), endothelin-1 (ET-1) and intercellular adhesion molecule-1 (ICAM-1) of ECV304 cells incubated with HG was increased obviously, but ECV304/IκBαM cells could resist HG induced increase of the expression of inflammatory factors. Conclusions HG increase the expression of inflammatory factors in vascular endothelial cells. Inhibition of nuclear factor-κB activation can protect vascular endothelial cells from the cytotoxicity of HG.